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In the healthy primate, neurons of the external and internal segments of the globus pallidus GP present a primarily irregular firing pattern, and a negligible level of synchrony is observed between pairs of neurons. This holds even for neighboring cells, despite their higher probability to receive common inputs and to innervate each other via lateral connectivity. In the Parkinsonian primate, this changes drastically, and many pairs of GP cells show synchronous oscillations. To address the relation between distance and synchrony in the Parkinsonian state, we compared the synchrony of discharge of close pairs of neurons, recorded by the same electrode, with remote pairs, recorded by different ones. However, spike trains of neighboring cells recorded by the same extracellular electrode exhibit the shadowing effect; i.e., lack of detection of spikes that occur within a few milliseconds of each other. Here, we demonstrate that the shadowing artifact can both induce apparent correlations between non-correlated neurons, as well as conceal existing correlations between neighboring ones. We therefore introduced artificial shadowing in the remote pairs, similar to the effect we observed in the close ones. After the artificial shadowing, neighboring cells did not show a higher tendency to oscillate synchronously than remote ones. On the contrary, the average percentage over all sessions of artificially shadowed remote pairs exhibiting synchronous oscillations was 35.4% compared to 17.2% in the close ones. A similar trend was found when the unshadowed remote pairs were separated according to the estimated distance between electrode tips: 29.9% of pairs at approximate distance of less than 750 m were significantly synchronized, in comparison with 28.5% of the pairs whose distance was more than 750 m. We conclude that the synchronous oscillations in the GP of MPTP-treated primates are homogenously distributed. The primary goal of Bio-Quick is to provide a vehicle to transform the innovative biotechnology available in research labs from Armed Forces Institute of Pathology into marketable and profitable medical instruments that can greatly benefit the health care, food market safety control, and the advancement in medical community. Formalin-fixation and paraffin-embedding FFPE is a time consuming but standard tissue preservation and processing method used in over 90 percent cases in hospitals and clinical settings for routine histology diagnosis. Our proposed project is to design and develop an ultrasound-facilitated processor DTP for rapid tissue fixation and processing for histology diagnosis and any further molecular study if necessary. The implementation of the technique will allow a significant reduction in processing time from at least 24 hours by conventional FFPE to less than 1 hour. We also need funding to support collaboration with outside and independent researchers to provide objective evaluation of the technique. Our specific aims of this SBIR phase I project are: 1 Development of a commercialized intensity adjustable bench top fixer/processor for rapid formalin fixation and paraffin embedding, 2 Evaluate and validate the DTP method in comparison to conventional FFPE method based on preservation of morphological details and molecular analyses. During the past 7 years, we have compared the DTP method with conventional FFPE method on over 100 human tissue specimens of 14 tissue types. Our preliminary data have demonstrated that compared to conventional FFPE, US-facilitated FFPE not only significantly reduces the total fixation/processing time from over 24 hours to within 1 hour, but also preserves similar or better tissue morphology, much improved protein antigen properties and mRNA integrity. As a result of improved preservation of macromolecules, antigen retrieval treatment prior to IHC staining may be reduced, much reduced 20X or more antibody concentration and shortened IHC reaction time are used. Long term stability of tissue morphology and mRNA integrity in USFFPE tissues is slightly better than that in conventional FFPE tissues.

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Posted by at June 9, 2015
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